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RNA interference of the salivary gland nitrophorin 2 in the triatomine bug Rhodnius prolixus (Hemiptera : Reduviidae) by dsRNA ingestion or injection

Araujo, R. N., Santos, A., Pinto, F. S., Gontijo, N. F., Lehane, Mike and Pereira, M. H. (2006) 'RNA interference of the salivary gland nitrophorin 2 in the triatomine bug Rhodnius prolixus (Hemiptera : Reduviidae) by dsRNA ingestion or injection'. Insect Biochemistry and Molecular Biology, Vol 36, Issue 9, pp. 683-693.

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Abstract

Mass sequencing of cDNA libraries from salivary glands of triatomines has resulted in the identification of many novel genes of unknown function. The aim of the present work was to develop a functional RNA interference (RNAi) technique for Rhodnius prolixus, which could be widely used for functional genomics studies in triatomine bugs. To this end, we investigated whether double-stranded RNA (dsRNA) can inhibit gene expression of R. prolixus salivary nitrophorin 2 (NP2) and what impact this might have on anticoagulant and apyrase activity in the saliva. dsRNA was introduced by two injections or by ingestion. RT-PCR of the salivary glands showed that injections of 15 mu g of NP2 dsRNA in fourth-instar nymphs reduced gene expression by 75 +/- 14% and that feeding 1 mu g/mu L of NP2 dsRNA into second-instar nymphs (approx. 13 mu g in total) reduced gene expression by 42 +/- 10%. Phenotype analysis showed that saliva of normal bugs prolonged plasma coagulation by about four-fold when compared to saliva of knockdown bugs. These results and the light color of the salivary gland content from some insects are consistent with the knockdown findings. The findings suggest that RNAi will prove a highly valuable functional genomics technique in triatomine bugs. The finding that feeding dsRNA can induce knockdown is novel for insects. (c) 2006 Elsevier Ltd. All rights reserved.

Item Type: Article
Uncontrolled Keywords: triatomine rhodnius prolixus rna interference nitrophorin salivary gland double-stranded-rna functional genomic analysis genetic interference bloodsucking insect host immunoglobulin passage anopheles antibody elegans protein
Subjects: QU Biochemistry > QU 4 General works
QU Biochemistry > Genetics > QU 450 General Works
QX Parasitology > QX 20 Research (General)
QX Parasitology > QX 4 General works
QX Parasitology > Insects. Other Parasites > QX 503 Hemiptera (e.g., Bedbugs)
Faculty: Department: Groups (2002 - 2012) > Vector Group
Digital Object Identifer (DOI): https://doi.org/10.1016/j.ibmb.2006.05.012
Depositing User: Sarah Lewis-Newton
Date Deposited: 21 Feb 2011 15:04
Last Modified: 06 Feb 2018 13:01
URI: http://archive.lstmed.ac.uk/id/eprint/1441

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